Amino Acid distance into crystallized protein?

Hello, Is someone know if it's possible from a crystallized protein modelisation to know the distance between X and Y amino acid? Which softwares/ websites to consider? Thanks a lot in...

05 June 2019 8,272 4 View

Detection of PAK proteins in HL60 cell line using western-blot?

Hello, I am trying to detect PAK1, PAK2 in cell lysates from HL60 cell line in western-blot. I tried to go for 50ug of protein but I just had a very tiny little band. I thought 50 ug of prot...

06 July 2018 9,233 0 View

NIH3T3 in Soft agar assay, max time of incubation?

Hi all, I am using the CellBioLabs kit to do my soft agar assay. I am quite surprised to see in the protocol the time incubation for NIH3T3-Ras transfected cells is 8 days? It seems to me quite...

06 July 2016 4,751 2 View

Ras G12V mutant issue in Soft agar assay?

Hi All, I am starting to do some soft agar assays using NIH/3T3 cells. As positive control I used NIH transfected with RasG12V plasmid. Before placing my cell on the agar gel I noticed (twice and...

06 July 2016 9,761 1 View

Does anyone has expereinces in NIH 3T3 electroporation using NEON system fron Invitrogen?

05 June 2016 7,330 2 View

Jurkat activation by LPS?

05 June 2016 7,847 2 View

What does Servant Leadership mean today and in the future? What are the advantages and disadvantages?

In current time of many changes in business and world of work, a lot is said and reported about the importance, skills and benefits of leadership. Are soft skills becoming more and more important?...

02 March 2021 8,149 3 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 2 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View